Why cells don't adhere well

Keeping cells well is the premise of scientific research. The cell culture procedure seems simple, but the slightest mistake can trigger a "do-over" mechanism, cell non-adherence is a microcosm of many problems.

Today we're going to talk about：

Why aren't your cells adhering to the wall?

How do promote cells to better adhere to the wall?

What is associated with cell adhesion?

First of all, not all cells will adhere when cultured in vitro, but most cells from solid tissues and organs do.

Cell adhesion process: Cells first secrete an extracellular matrix, which adheres to the surface of the support, and cells bind to these extracellular matrices through adhesion factors expressed on the surface.

Therefore, whether or not cells adhere to the cell wall is related to the ability of the cell itself to secrete extracellular matrix and the number of adhesion molecules expressed by the cell itself, as well as the surface structure of the culture vessel.

So~~~here is the point

Analysis of the reasons why cells do not adhere to the wall

1. Improper control of the trypsin digestion time, that means if the digestion is not enough, the cells themselves will form agglomerates; If the trypsin treatment is too long, it is easy to cause damage to the cell membrane proteins, so that the cells are not tightly adhered to the wall, and the stereoscopic effect under the microscope is not strong, and even in severe cases. cause cell death.

2. Lack of adhesion-promoting factors: Serum contains adhesion-promoting factors, and the lack of such adhesion-promoting factors in the serum-free medium process will greatly reduce the adherence effect of cells.

3. Mycoplasma or bacterial contamination.

4. The recovered cells are in poor condition, have aged, and lost their adhesion.

5. The number of seeded cells is too small to secrete a sufficient extracellular matrix.

Do like this to promote cell adhesion

1. Moderate digestion of cells;

2. It is recommended to use surface-treated disposable plastic containers for culture;

3. Use appropriate extracellular matrix or adherent reagents;

4. To revive new cells, use 20% serum to help cells recover in the first week;

5. The subculture and inoculation must be spread evenly to avoid cell clumping growth;

6.?Within 24 hours of cell passage, try not to shake, so as not to affect the adherence.

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